Skip to main content Accessibility help

Viability and apoptosis in spermatozoa of transgenic rabbits

  • P. Chrenek (a1) (a2), A.V. Makarevich (a3) and M. Simon (a4)


The aim of our study was to compare the viability of sperm cells from transgenic (mWAP-hFVIII gene) or non-transgenic (normal) rabbit males as assessed by viability (SYBR-14/PI) and apoptosis (annexin V) tests. These results were evaluated using female conception rates following insemination with the respective sperm samples. No significant differences were found in concentration and motility between transgenic and non-transgenic spermatozoa. Spermatozoa from both transgenic (63.05 ± 20.05%) or non-transgenic (65.75 ± 22.15%) males, stained with SYBR-14 (green), were found to be morphologically normal. In both groups, the highest proportion of annexin V-positive sperm staining was found in the post-acrosomal part of the sperm head (8.66 and 27.53%). The percentage of sperm that stained with SYBR-14/PI or with annexin V/DAPI was correlated with liveborn in transgenic rabbits (R2 = 0.6118 and R2 = 0.2187, respectively) or non-transgenic rabbits (R2 = 0.671 and R2 = 0.3579, respectively). These data indicate that there was no difference in the viability of rabbit transgenic and non-transgenic spermatozoa when determined by both fluorescence assays.


Corresponding author

All correspondence to: P. Chrenek. Animal Production Research Centre Nitra, 95141 Luzianky near Nitra, Slovak Republic. Tel: + 421 37 6546 289. Fax: + 421 37 6546 285. e-mail:


Hide All
Bialkowska, J., Demianowicz, W. & Glogowski, J. (2004). Determining the viability of zebra mussel (Dreissena polymorpha) spermatozoa and changes in the integrity of its membrane using the fluorescence method. Arch. Polish Fish. 12, 2330.
Chrenek, P., Vasicek, D., Makarevich, A., Jurcik, R., Suvegova, K., Bauer, M., Parkanyi, V., Rafay, J., Batorova, A. & Paleyanda, R.K. (2005). Increased transgene integration efficiency upon microinjection of DNA into both pronuclei of rabbit embryos. Transgenic Res. 14, 417–28.
Chrenek, P., Dragin, S. & Makarevich, A.V. (2006). Reproductive characteristics of transgenic rabbit males with human protein C gene. Slovak J. Anim. Sci. 39, 131–4.
Chrenek, P., Trandzik, J., Massanyi, P., Makarevich, A., Lukac, N., Peskovicova, D. & Paleyanda, R.K. (2007a). Effect of transgenesis on reproductive traits of rabbit males. Anim. Reprod. Sci. 99, 127–34.
Chrenek, P., Massányi, P., Makarevich, A.V., Pukač, N., Zahradnikova, M., Schneidgenova, M. & Ryban, L. (2007b). Rabbit transgenic brothers with different reproductive traits. Slovak J. Anim. Sci. 40, 113–7.
Gadella, B.M. & Harrison, R.A.P. (2002). Capacitation induces cyclic adenosine 3′,5′-monophosphate-dependent, but apoptosis-unrelated, exposure of aminophospholipids at the apical head plasma membrane of boar sperm cells. Biol. Reprod. 67, 1340–50.
Garner, D.L. & Johnson, L. (1995). Viability assessment of mammalian spermatozoa using SYBR-14 and propidium iodide. Biol. Reprod. 53, 276–84.
Garner, D.L., Johnson, L.A., Allen, C.H., Palencia, D.D. & Chambers, C.S. (1996). Comparison of seminal quality in Holstein bull as yearling and as mature sires. Theriogenology 45, 923–34.
Harrison, R.A. & Vickers, S.E. (1990). Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. J. Reprod. Fertil. 88, 343–52.
Hong, C.Y., Juany, J.J., Wu, P., Lo, S. & Wei, Y.H. (1988). Fluorescence supravita stain human sperm correlation with sperm motility measured by a transmembrane migration metod. Andrologia 20, 516–20.
Januskauskas, A., Johannisson, A. & Rodriguez-Martinez, H. (2003). Sibyle membrane changes in cryopreserved bull semen in relation with sperm viability chromatin structure, and field fertility. Theriogenology 60, 743–58.
Makarevich, A.V., Parkanyi, V., Ondruška, Ľ., Kubovičová, E., Fľak, P., Slezáková, M., Pivko, J. & Rafay, J. (2008a). Evaluation of fertilizing capacity of rabbit sperm on the basis of annexin V-labelled membrane changes. Slovak J. Anim. Sci. 41, 15.
Makarevich, A.V., Chrenek, P., Massányi, P., Lukac, N. & Pivko, J. (2008b). Apoptosis as a tool for determination of animal embryo quality. Slovak J. Anim. Sci. 41, 153–9.
Martin, G., Sabido, O., Durand, P. & Levy, R. (2004). Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol. Reprod. 71, 28–37.
Muratori, M., Porazzi, I., Luconi, M., Marchiani, S., Forti, G. & Baldi, E. (2004). Annexin V binding and merocyanine staining fail to detect human sperm capacitation. J. Androl. 25, 797810.
Pena, F.J., Johannisson, A., Wallgren, M. & Rodriguez-Martinez, H. (2003). Assessment of fresh and frozen–thawed boar semen using an annexin-V assay: a new method of evaluating sperm membrane integrity. Theriogenology 60, 677–89.
Vermes, I., Haanen, C., Steffens-Nakken, H. & Reutelingsperger, C.P.M. (1995). A novel assay for apoptosis: flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J. Immunol. Meth. 184, 3951.
Vetter, C.M., Millet, J.E., Crawford, L.M., Armstrong, M.J., Clair, J. H., Conner, M.W., Wise, L.D. & Skopek, T.R. (1998). Comparison motility and membrane integrity to assess sperm viability. Reprod. Toxicol. 12, 105–14.


Related content

Powered by UNSILO

Viability and apoptosis in spermatozoa of transgenic rabbits

  • P. Chrenek (a1) (a2), A.V. Makarevich (a3) and M. Simon (a4)


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.